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Research Project: Integrated Research to Improve On-Farm Animal Health in Salmonid Aquaculture

Location: Cool and Cold Water Aquaculture Research

Title: Shotgun proteomic analysis of Yersinia ruckeri isolates under normal and iron-limited conditions

item Kumar, Gokhlesh - University Of Veterinary Medicine
item Hummel, Karin - University Of Veterinary Medicine
item Ahrens, Maike - Ruhr-University Bochum
item Menanteau-ledouble, Simon - University Of Veterinary Medicine
item Razzazi-fazeli, Ebrahim - University Of Veterinary Medicine
item Welch, Timothy - Tim
item El-matbouli, Mansour - University Of Veterinary Medicine

Submitted to: Veterinary Research
Publication Type: Peer Reviewed Journal
Publication Acceptance Date: 9/9/2016
Publication Date: 10/6/2016
Citation: Kumar, G., Hummel, K., Ahrens, M., Menanteau-Ledouble, S., Razzazi-Fazeli, E., Welch, T.J., El-Matbouli, M. 2016. Shotgun proteomic analysis of Yersinia ruckeri isolates under normal and iron-limited conditions. Veterinary Research. 47:100. doi:10.1186/s13567-016-0384-3.

Interpretive Summary: Bacteria differentially regulate proteins in order to adapt to changes in their environment, including adaptation to the conditions that pathogens face within their host during the infection process. Host sequestration of iron is known to be a critical innate defence strategy as limiting pathogen access to this essential micronutrient restricts bacterial growth during infection. In this study, we used proteomic analysis to determine the whole cell protein expression pattern of the fish pathogen Yersinia ruckeri grown in iron replete and iron limited conditions. Sixty one proteins were found to be regulated by iron availability in Y. ruckeri. These proteins were involved in iron scavenging and transport as well as other metabolic processes. Differential regulation was confirmed by assessing transcriptional regulation of the genes encoding these proteins using quantitative real time PCR. This paper presents the first analysis of the Y. ruckeri proteome and provides a better understanding of the proteins expressed under the iron restrictive conditions within the host during infection.

Technical Abstract: Yersinia ruckeri is the causative agent of enteric redmouth disease of fish and causes significant economic losses, particularly in salmonids. Iron is an essential nutrient for many cellular processes and is involved in host sensing and virulence regulation in many bacteria. Bacterial pathogens differentially express proteins in the host during the infection process or under certain environmental conditions. Little is known about proteomics expression of Y. ruckeri in response to iron-limited conditions. Here, we present whole cell protein identification and quantification for two motile and two non-motile isolates of Y. ruckeri under iron-sufficient and iron-limited conditions using a shotgun proteomic approach. Label-free gel-free quantification was performed by using a nanoLC-ESI and high resolution mass spectrometry. SWATH technology was applied in this approach in order to distinguish between different isolates and their response to iron withdrawal. A total of 61 differentially expressed proteins were identified in four Y. ruckeri isolates under iron-limited conditions. The identified proteins were involved in various processes, including iron ion capture and transport as well as enzymatic metabolism. The proteins identified by shotgun proteomic approach were confirmed to be differentially expressed at the transcriptional level using quantitative real time PCR. Upregulation of iron-restricted proteins suggests their involvement in the pathogenesis of Y. ruckeri. Our study provides the first detailed proteome analysis of Y. ruckeri isolates and will contribute to understanding virulence mechanisms of Y. ruckeri as well as the development of novel control methods for enteric redmouth disease.